IGEM:Peking University/2008/Experiment/PKU 0807018 G1 1: Difference between revisions
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::'''Gel running-testing-pADH-lac1/S/tet''' | |||
::'''Gel running-testing-digestion-lac2/AID''' | |||
Revision as of 15:21, 18 July 2008
PKU_080718_G1_1
- Enzyme cutting-pADH-lac2(PCR product)
- System for double ligation
| Volume (μL) | |
| Promoter | 20 |
| BamHI | 1 |
| SacI | 1 |
| 0.5×K Buffer | 2.5 |
| ddH2O | 25.5 |
- System for single ligation
| Volume (μL) | |
| Promoter | 20 |
| XhoI | 2 |
| SacI | 2 |
| 1×M Buffer | 5 |
| ddH2O | 21 |
- 'PCR- pGREG504-pADH-(lac1/lacS/tet)-50ul
- Using program lac12MQT
- Gel running and extraction-promoter(PCR product)
- Enzyme cutting-AID
- lanes: intact AID(control)|λmarker|Promoter(single)|promoter(double)|AID|AID
- System for double ligation
| Volume (μL) | ||
| Promoter | 20 | |
| BamHI | 1 | ×3 |
| SacI | 1 | |
| 0.5×K Buffer | 2.5 | |
| ddH2O | 25.5 |
- System for single ligation
| Volume (μL) | ||
| Promoter | 20 | |
| XhoI | 2 | ×3 |
| SacI | 2 | |
| 1×M Buffer | 5 | |
| ddH2O | 21 |
- Single Ligation-pGREG504-pAID-lac2
| Volume (μL) | |
| lac2 | 4 |
| pGREG504 | 1 |
| ligation buffer | 5 |
- Double Ligation-pGREG504-pAID-lac2-AID
| Volume (μL) | |
| lac2 | 3 |
| AID | 3 |
| pGREG504 | 1 |
| ligation buffer | 7 |
- Gel running-testing-pADH-lac1/S/tet
- Gel running-testing-digestion-lac2/AID
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